Optimisation of a Capillary Zone Electrophoresis Methodology for Simultaneous Analysis of Organic Aliphatic Acids in Extracts of Brachiaria brizantha
Marcone Augusto Leal de Oliveira, Department of Chemistry, Federal University of Juiz de Fora - UFJF, 36036–330, Juiz de Fora, MG, Brazil. E-mail: email@example.com
Aluminum toxicity is commonly verified in acidic soils, and poses a severe limitation to plant growth and development. Therefore, Al complexation by the root system mucilage, Al complexation by organic compounds that are exuded by the roots and internal metabolic processes must be monitored by organic acids (OA), since they play a central role in these aluminum tolerance mechanisms.
To optimise a capillary zone electrophoresis method able to perform simultaneous separation of acetic, citric, formic, lactic, malic, oxalic, pyruvic, succinic, tartaric and aspartic acid in plant extract solutions.
Method optimisation was achieved by a chemometric approach through experimental designs. The optimal condition found was: 20 mmol/L phthalic acid buffer; 0.8 mmol/L cetyltrimethyl-ammonium bromide; pH 3.4 adjusted with tris(hydroxymethyl)aminomethane (around 16 mmol/L); −15 kV of voltage; 25 °C of cartridge temperature; indirect ultraviolet detection at 240 nm; and 25 mbar injection for 2 s, within an analysis time of 4 min.
As a repeatability test of the optimal condition, 30 replicates were carried out with the same working electrolyte, where the relative standard deviation of each peak ranged from 0.081 to 0.36% (for migration time) and from 2.4 to 4.6% (for peak area).
The methodology was successfully applied to simultaneously determine citric, malic and aspartic acid in roots and leaves extract solutions of Brachiaria brizantha, demonstrating its usefulness to study aluminum tolerance. Copyright © 2012 John Wiley & Sons, Ltd.